Chicken-derived cd20 antibodies with potent b cell depletion activity

Abstract

The present disclosure provides multiple anti-hCD20 mAbs as well as humanization of antibodies. The antibodies described herein are chicken-derived and exhibit significantly enhanced B-cell-specific antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) potency as well as improved depletion of B lymphoma cells in vivo relative to Rituximab.

1 . An antigen-binding molecule that specifically binds to CD20, wherein the antigen-binding molecule comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein the VH comprises a complementarity determining region 1 (VH CDR1) comprising the amino acid sequence of SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 13 and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 17; and wherein the VL comprises a complementarity determining region 1 (VL CDR1) comprising the amino acid sequence of SEQ ID NO: 21, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 25, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 29: 
       
         
           
                 
                 
               
                     
                   VH CDR1 
                 
                     
                   (SEQ ID NO: 9) 
                 
                     
                   SSYAM 
                 
                     
                     
                 
                     
                   VH CDR2 
                 
                     
                   (SEQ ID NO: 13) 
                 
                     
                   EITNYAAGSGTWYGAAVK 
                 
                     
                     
                 
                     
                   VH CDR3 
                 
                     
                   (SEQ ID NO: 17) 
                 
                     
                   KGPISGSIGYLSSIDA 
                 
                     
                     
                 
                     
                   VL CDR1 
                 
                     
                   (SEQ ID NO: 21) 
                 
                     
                   SGGSSRYGYG 
                 
                     
                     
                 
                     
                   VL CDR2 
                 
                     
                   (SEQ ID NO: 25) 
                 
                     
                   WNDKRPS 
                 
                     
                     
                 
                     
                   VL CDR3 
                 
                     
                   (SEQ ID NO: 29) 
                 
                     
                   GNYDNNDPA. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The antigen-binding molecule of  claim 1 , wherein the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1. 
     
     
         3 .- 13 . (canceled) 
     
     
         14 . The antigen-binding molecule of  claim 1 , wherein the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 5. 
     
     
         15 .- 25 . (canceled) 
     
     
         26 . An antigen-binding molecule that specifically binds to CD20, wherein the antigen-binding molecule comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein the VH comprises a complementarity determining region 1 (VH CDR1) comprising the amino acid sequence of SEQ ID NO: 10, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 14 and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 18; and wherein the VL comprises a complementarity determining region 1 (VL CDR1) comprising the amino acid sequence of SEQ ID NO: 22, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 26, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 30: 
       
         
           
                 
                 
               
                     
                   VH CDR1 
                 
                     
                   (SEQ ID NO: 10) 
                 
                     
                   SSSYIN 
                 
                     
                     
                 
                     
                   VH CDR2 
                 
                     
                   (SEQ ID NO: 14) 
                 
                     
                   QINKDGGGSWYAPVVK 
                 
                     
                     
                 
                     
                   VH CDR3 
                 
                     
                   (SEQ ID NO: 18) 
                 
                     
                   KNADSGCVGVGGCIDT 
                 
                     
                     
                 
                     
                   VL CDR1 
                 
                     
                   (SEQ ID NO: 22) 
                 
                     
                   SGGSYYYGGNYYYG 
                 
                     
                     
                 
                     
                   VL CDR2 
                 
                     
                   (SEQ ID NO: 26) 
                 
                     
                   NNNKRPS 
                 
                     
                     
                 
                     
                   VL CDR3 
                 
                     
                   (SEQ ID NO: 30) 
                 
                     
                   GGYDSSYVAI. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         27 . The antigen-binding molecule of  claim 26 , wherein the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. 
     
     
         28 .- 38 . (canceled) 
     
     
         39 . The antigen-binding molecule of  claim 26 , wherein the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 6. 
     
     
         40 .- 50 . (canceled) 
     
     
         51 . The antigen-binding molecule of  claim 26 , wherein the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 33. 
     
     
         52 .- 62 . (canceled) 
     
     
         63 . The antigen-binding molecule of  claim 26 , wherein the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 34. 
     
     
         64 .- 74 . (canceled) 
     
     
         75 . An antigen-binding molecule that specifically binds to CD20, wherein the antigen-binding molecule comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL), wherein the VH comprises a complementarity determining region 1 (VH CDR1) comprising the amino acid sequence of SEQ ID NO: 11, a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 15 and a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 19; and wherein the VL comprises a complementarity determining region 1 (VL CDR1) comprising the amino acid sequence of SEQ ID NO: 23, a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 31: 
       
         
           
                 
                 
               
                     
                   VH CDR1 
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   NSNGMG 
                 
                     
                     
                 
                     
                   VH CDR2 
                 
                     
                   (SEQ ID NO: 15) 
                 
                     
                   GIHSSGRYTYYGTAVK 
                 
                     
                     
                 
                     
                   VH CDR3 
                 
                     
                   (SEQ ID NO: 19) 
                 
                     
                   KNADSAYGYWYAGSIDA 
                 
                     
                     
                 
                     
                   VL CDR1 
                 
                     
                   (SEQ ID NO: 23) 
                 
                     
                   SGGYSSYGYS 
                 
                     
                     
                 
                     
                   VL CDR2 
                 
                     
                   (SEQ ID NO: 27) 
                 
                     
                   NNNNRPS 
                 
                     
                     
                 
                     
                   VL CDR3 
                 
                     
                   (SEQ ID NO: 31) 
                 
                     
                   AFTDYSSLAGV. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         76 . The antigen-binding molecule of  claim 75 , wherein the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 3. 
     
     
         77 .- 87 . (canceled) 
     
     
         88 . The antigen-binding molecule of  claim 75 , wherein the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 7. 
     
     
         89 .- 124 . (canceled) 
     
     
         125 . The antigen-binding molecule of  claim 1 , wherein the antigen-binding molecule is an antibody or a CD20-binding fragment thereof. 
     
     
         126 . The antigen-binding molecule of  claim 125 , wherein the CD20-binding fragment is selected from the group consisting of an Fab fragment, an scFab, an Fab′, a single chain variable fragment (scFv) and a one-armed antibody. 
     
     
         127 . The antigen-binding molecule of  claim 1 , wherein the antigen-binding molecule is a humanized antibody or CD20-binding fragment thereof. 
     
     
         128 .- 146 . (canceled) 
     
     
         147 . The antigen-binding molecule of  claim 26 , wherein the antigen-binding molecule is an antibody or a CD20-binding fragment thereof. 
     
     
         148 . The antigen-binding molecule of  claim 147 , wherein the CD20-binding fragment is selected from the group consisting of an Fab fragment, an scFab, an Fab′, a single chain variable fragment (scFv) and a one-armed antibody. 
     
     
         149 . The antigen-binding molecule of  claim 26 , wherein the antigen-binding molecule is a humanized antibody or CD20-binding fragment thereof. 
     
     
         150 . The antigen-binding molecule of  claim 75 , wherein the antigen-binding molecule is an antibody or a CD20-binding fragment thereof. 
     
     
         151 . The antigen-binding molecule of  claim 150 , wherein the CD20-binding fragment is selected from the group consisting of an Fab fragment, an scFab, an Fab′, a single chain variable fragment (scFv) and a one-armed antibody. 
     
     
         152 . The antigen-binding molecule of  claim 75 , wherein the antigen-binding molecule is a humanized antibody or CD20-binding fragment thereof.